Bazı Üzüm Çeşitlerinde (Vitis vinifera L.) N2O Uygulamalarıyla Ototetraploidi Teşviki
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Dosyalar
Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Doğal veya kontrollü uyarıcıların etkisiyle poliploid bitkiler oluşabilmekte olup bu bitkiler farklı
çevresel streslerle başa çıkma ve tüketicilerin talep ettiği ürün kalitesini sağlamada bir araç olabildiğinden,
son zamanlarda asma ıslahında dikkat çekmektedir. Global düzeyde poliploid üzüm çeşitlerine ilginin ve
özellikle bu çeşitlerin bağ alanlarının çok hızlı artarak tüm üzüm çeşitleri arasında ilk sıraya ulaşması üzüm
çeşitlerinin poliploidi yöntemleriyle geliştirilmesi çabalarını artırmıştır. Bu çalışmada, bitkisel materyal
olarak, +4 °C’de 3 ay katlanan Gök Üzüm, Ekşi Kara, Trakya İlkeren ve Narince üzüm çeşitlerinin (Vitis
vinifera L.) tohumları ve odun çelikleri poliploidi uyarımı için, 24, 48 ve 72 saat 2.5, 5, 7.5 ve 10 bar N2O
dozlarına maruz bırakıldı. N2O’nun materyallerde canlılık oranı (%), ölümcül doz (LD), stoma
parametreleri (en, boy, yoğunluk, kloroplast sayısı), klorofil miktarı (SPAD değeri), nDNA içeriği ve
kromozom sayılarına etkileri incelendi. Stoma gözlemleri, klorofil ve kloroplast sayımlarıyla seçilmiş
örneklerde flow sitometri (FC) analizi ve analog koromozom sayımıyla ploidi doğrulamaları yapıldı.
Mutagen uygulanan materyalin canlılık oranlarında ortaya çıkan farklılıklara, iklim etkileriyle genotiplere
göre değişen tomurcuk nekrozu ve N2O’ya tepkideki genotipik farklılıklarının neden olduğu düşünüldü.
Ölümcül doz hesaplamalarında N2O etkilerinin doğrusal olmadığı, parabolik bir değişim sergilediği
belirlendi. N2O tüm asma materyallerinde stoma boyu, stoma yoğunluğu, kloroplast sayısı ve klorofil
miktarlarını önemli (p<0.05) düzeyde etkiledi. Stoma boyutundaki değişim nedeniyle mutasyon oluştuğu
varsayılan materyallerde FC analizleriyle doğrulama testleri yapıldı. FC analizleriyle 7.5 bar N2O ile 24
saat muamele edilen bir Gök Üzüm fidesinin triploid/yakın triploid olduğu tespit edildi. Seçilen genotipte
nDNA içeriğinin 1.57 pg/2C olduğu belirlendi ve 1C genom boyutu 768 Mbp olarak hesaplandı. Seçilmiş
poliploid genotipin yaprak örneğinde kromozom sayımı yapıldı ve görüntüleri alındı. Stoma gözlemleri,
FC analizleri ve kromozom sayımı sonuçları, asma tohumlarında kromozom katlanmasında N2O’nun etkili
olduğunu gösterdi. Bundan sonraki asmada poliploidi uyarımına yönelik çalışmalarda farklı çeşitler için
N2O’nun bir mutagen olarak kullanılabileceği, daha fazla poliploid asma genotipleri elde etmek için 7.5 bar
24 saat uygulamasının yanı sıra farklı doz ve uygulama sürelerinin denenmesinin uygun olacağı
düşünülmektedir.
Polyploid plants can be formed under the influence of natural or controlled stimulants and have recently attracted attention in grapevine breeding as they can be a means of coping with different environmental stresses and providing the product quality demanded by consumers. The global interest in polyploid grape varieties, and especially the rapid increase in the vineyard areas of these varieties and reaching the first place among all grape cultivars, has increased the efforts to develop grape varieties by polyploid methods. In this study, as plant material, seeds, and hard wood cuttings of Gök Üzüm, Ekşi Kara, Trakya İlkeren and Narince grape varieties (Vitis vinifera L.) cold-stratified at +4 °C for 3 months were used for polyploidy induction and they exposed to 2.5, 5, 7.5 and 10 bar N2O doses for 24, 48 and 72 hours. The effects of N2O on the survival rate (%), lethal dose (LD), stoma parameters (width, length, density, chloroplast number), chlorophyll amount (SPAD value), nDNA content and chromosome numbers were determined. Ploidy confirmations were performed in selected samples with stoma observations, chlorophyll, and chloroplast counts by flow cytometry (FC) analysis and analogue chromosome counting. It was thought that the differences in the viability of the material applied mutagen were caused by the differences in bud necrosis and N2O response, which varies according to the genotypes with climatic effects. In lethal dose calculations, it was determined that the effects of N2O were non-linear and showed a parabolic change. N2O significantly (p<0.05) affected stomatal length, stomatal density, chloroplast number and chlorophyll amount in all grapevine materials. FC analyses were performed on materials assumed to have mutations due to the change in stoma size. From these materials, it was determined by FC analysis that a Gök Üzüm seedling obtained by treating the seeds with 7.5 bar N2O for 24 hours was triploid/near triploid genotype. The nDNA content of the selected genotype was determined as 1.57 pg/2C and the 1C genome size was calculated as 768 Mbp. Chromosome images were obtained from leaf samples of the selected polyploid genotype. Stoma observations, FC analyses, and chromosome number observation results showed that N2O was effective in chromosome doubling for grapevine seeds. It is thought that N2O can be used as a mutagen for different varieties in the future studies on induction of polyploidy in grapevine, and it would be appropriate to try different doses and application times as well as 7.5 bar 24 hours application to obtain more polyploid grapevine genotypes.
Polyploid plants can be formed under the influence of natural or controlled stimulants and have recently attracted attention in grapevine breeding as they can be a means of coping with different environmental stresses and providing the product quality demanded by consumers. The global interest in polyploid grape varieties, and especially the rapid increase in the vineyard areas of these varieties and reaching the first place among all grape cultivars, has increased the efforts to develop grape varieties by polyploid methods. In this study, as plant material, seeds, and hard wood cuttings of Gök Üzüm, Ekşi Kara, Trakya İlkeren and Narince grape varieties (Vitis vinifera L.) cold-stratified at +4 °C for 3 months were used for polyploidy induction and they exposed to 2.5, 5, 7.5 and 10 bar N2O doses for 24, 48 and 72 hours. The effects of N2O on the survival rate (%), lethal dose (LD), stoma parameters (width, length, density, chloroplast number), chlorophyll amount (SPAD value), nDNA content and chromosome numbers were determined. Ploidy confirmations were performed in selected samples with stoma observations, chlorophyll, and chloroplast counts by flow cytometry (FC) analysis and analogue chromosome counting. It was thought that the differences in the viability of the material applied mutagen were caused by the differences in bud necrosis and N2O response, which varies according to the genotypes with climatic effects. In lethal dose calculations, it was determined that the effects of N2O were non-linear and showed a parabolic change. N2O significantly (p<0.05) affected stomatal length, stomatal density, chloroplast number and chlorophyll amount in all grapevine materials. FC analyses were performed on materials assumed to have mutations due to the change in stoma size. From these materials, it was determined by FC analysis that a Gök Üzüm seedling obtained by treating the seeds with 7.5 bar N2O for 24 hours was triploid/near triploid genotype. The nDNA content of the selected genotype was determined as 1.57 pg/2C and the 1C genome size was calculated as 768 Mbp. Chromosome images were obtained from leaf samples of the selected polyploid genotype. Stoma observations, FC analyses, and chromosome number observation results showed that N2O was effective in chromosome doubling for grapevine seeds. It is thought that N2O can be used as a mutagen for different varieties in the future studies on induction of polyploidy in grapevine, and it would be appropriate to try different doses and application times as well as 7.5 bar 24 hours application to obtain more polyploid grapevine genotypes.
Açıklama
Anahtar Kelimeler
asma, flow sitometri, LD50, mitotik poliploidi, nDNA, N2O, ototetraploidi, Vitis vinifera L., grapevine, flow cytometry, mitotic polyploidy, autotetraploidy
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Özer, A., (2021). Bazı Üzüm Çeşitlerinde (Vitis vinifera L.) N2O Uygulamalarıyla Ototetraploidi Teşviki. (Doktora Tezi). Selçuk Üniversitesi, Fen Bilimleri Enstitüsü, Konya.
